Antisense oligonucleotides

ABSTRACT

The invention provides antisense DNA oligonucleotides which are effective in inhibiting the expression of a wild type COL 1 A 1  gene.

[0001] The present invention relates to antisense oligonucleotides and their use in inhibiting expression of type I procollagen.

[0002] The collagens are a family of closely related proteins, with a triple helix protein structure. Numerous collagen types have been identified (>10) of which type I procollagen (consisting of two alpha1 chains and one alpha2 chain) is the principal component of bone, skin, and tendon.

[0003] It has been recognised for many years that many pathological conditions are caused by overproduction of collagen fibres in the form of scars and excess fibrous tissues. For example, liver cirrhosis is a two-step process in which normal liver tissue is first destroyed by a virus or by alcohol and other toxins, and then excessive amounts of collagen fibres replace the damaged cells before normal liver cell regeneration. Idiopathic pulmonary fibrosis is a lethal condition in which normal lung tissue is gradually replaced by excessive amounts of collagen fibres. Progressive systemic sclerosis (scleroderma) is a frequently lethal disease where skin and many internal organs become leather-like because of excessive depositions of collagen fibres. In many individuals, wounds or surgical incisions in the skin are followed by excessive depositions of collagen in the form of hypertrophic scars and keloids that present cosmetic problems and sometimes more serious consequences. Also, excessive scarring frequently occurs in normal individuals following trauma and surgical procedures. In these and related conditions, a means of specifically inhibiting collagen synthesis and deposition would be of tremendous benefit.

[0004] PCT Patent Application Publication No. WO 94/11494 discloses a DNA or RNA oligonucleotide comprising from 5 to 200 nucleotides substantially complementary to a mutant collagen nucleotide sequence or a normal wild type collagen nucleotide sequence which is capable of inhibiting collagen gene expression. Preferred oligonucleotides are said to be antisense oligonucleotides. The Examples of WO 94/11494 describe a series of DNA oligonucleotides, some of which are antisense, that were synthesised primarily with regard to the region at the 3′ end of exon 1 (from nucleotides 198 to 222) and the first two nucleotides of intron 1 of the human gene for the proα1 chains of type I procollagen (COL1A1). The synthesised oligonucleotides were found to vary considerably in their ability to inhibit expression of an internally deleted mutant COL1A1 gene of human origin. The effectiveness of the oligonucleotides in inhibiting the expression of the human wild type COL1A1 gene was not however demonstrated. Since the structure and conformation of the RNA transcripts of the human, mutant and wild type COL1A1 genes would most likely differ, it would not necessarily follow that oligonucleotides which are effective inhibitors of the expression of the mutant COL1A1 gene would also be effective inhibitors of the expression of the wild type COL1A1 gene.

[0005] It would be desirable to identify antisense DNA oligonucleotides that are capable of inhibiting the expression of a wild type COL1A1 gene.

[0006] In accordance with the present invention, there is therefore provided an antisense DNA oligonucleotide comprising from 18 to 25 nucleotides which is complementary to a nucleotide sequence from position 750 to position 3900 inclusive of SEQ ID NO:1, wherein SEQ ID NO:1 comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence according to SEQ ID NO:2, the oligonucleotide being capable of inhibiting expression of the polypeptide in a cell that expresses it.

[0007] SEQ ID NO:1 is identical to the nucleotide sequence registered under EMBL accession no. Z74615. SEQ ID NO:2 is the amino acid sequence of the polypeptide encoded by the nucleotide sequence of SEQ ID NO:1. The polypeptide encoded by SEQ ID NO:1 is a precursor of the wild type, proα1 chain of type I procollagen (“prepro-alpha1 (I) collagen”).

[0008] The antisense DNA oligonucleotide according to the invention comprises 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides and is preferably 20 nucleotides in length.

[0009] The antisense DNA oligonucleotide is preferably complementary to a nucleotide sequence in one of the following regions of SEQ ID NO:1, Region 1 from position 750 to position 900 inclusive, Region 2 from position 1200 to position 1300 inclusive, Region 3 from position 1400 to position 1500 inclusive, Region 4 from position 1450 to position 1550 inclusive, Region 5 from position 1850 to position 2000 inclusive, Region 6 from position 2500 to position 2600 inclusive, Region 7 from position 2850 to position 2950 inclusive, Region 8 from position 3800 to position 3900 inclusive.

[0010] Particularly preferred antisense DNA oligonucleotides are those which are complementary to a nucleotide sequence in Region 2, 4, 6 or 8 of SEQ ID NO:1.

[0011] The oligonucleotides of the invention may be prepared by any suitable method known in the art. The oligonucleotides are very conveniently prepared by synthetic chemical methods, for example, phosphoramidite chemistry by sulfurization with tetraethylthiuram disulfide in acetonitrile as described in Tetrahedron Lett., 1991, 32, 30005-30008.

[0012] The oligonucleotides of the present invention are advantageous in that they inhibit expression of the wild type COL1A1 gene. They are therefore useful in the treatment or prevention of conditions/disorders caused by overproduction of collagen fibres, for example, liver cirrhosis, kidney, liver and heart fibrosis, scleroderma, hypertrophic scars and keloids.

[0013] Accordingly, the present invention provides an antisense DNA oligonucleotide according to the invention for use in therapy.

[0014] In another aspect, the invention provides the use of an antisense DNA oligonucleotide according to the invention in the manufacture of a medicament for use in therapy.

[0015] In the context of the present specification, the term “therapy” also includes “prophylaxis” unless there are specific indications to the contrary. The terms “therapeutic” and “therapeutically” should be construed accordingly.

[0016] The invention further provides a method of treating, or reducing the risk of, a collagen disorder in a patient suffering from, or at risk of, the disorder, which comprises administering to the patient a therapeutically effective amount of an antisense DNA oligonucleotide according to the invention.

[0017] For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the oligonucleotide employed, the mode of administration, the treatment desired and the disorder indicated. Effective dosages are those which are able to inhibit collagen protein production in cells at a level which eliminates or reduces the symptoms or conditions associated with the collagen protein production.

[0018] The oligonucleotides according to the invention will generally be administered in the form of a pharmaceutical composition in which the oligonucleotide is formulated with a pharmaceutically acceptable adjuvant, diluent or carrier.

[0019] Thus, the present invention also provides a pharmaceutical composition comprising an antisense DNA oligonucleotide according to the invention in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

[0020] The invention further provides a process for the preparation of a pharmaceutical composition of the invention which comprises mixing the antisense DNA oligonucleotide with a pharmaceutically acceptable adjuvant, diluent or carrier.

[0021] The pharmaceutical composition of the invention may be administered topically in the form of, for example, a creme, lotion or ointment; or systemically, e.g. by oral administration in the form of tablets, capsules, syrups, powders or granules, or by parenteral administration in the form of sterile solutions or suspensions.

[0022] The present invention will now be further explained by reference to the following illustrative Examples.

EXAMPLES Example 1

[0023] Oligonucleotide Synthesis

[0024] Phosphorothioate oligodeoxynucleotides synthesis was carried out at a 1 μm scale on PE Biosystems 394 DNA synthesizer using phosphoramidite chemistry with TETD/acetonitrile sulphurizing reagent. Oligonucleotides were purified on Poly-Pak TM II cartridges (Glen Research), desalted on NAPTM 10 columns (Amersham Pharmacia Biotech AB) and ion-exchanged using Dowex 50WX8-1100 ion exchange resin (Aldrich). Twelve antisense DNA oligonucleotides (ASOs) were prepared having the following sequences (5′→3′): 1. GGACGACCAGGTTTTCCAGC (SEQ ID NO:3) 2. GCACACCAGCAGGGCCAGG (SEQ ID NO:4) 3. GCCAGGAGCACCAGGTTCAC (SEQ ID NO:5) 4. CTTCCTCTCCAGCAGGGCCA (SEQ ID NO:6) 5. GCCTTGCCGGGCTCTCCAGC (SEQ ID NO:7) 6. CGGGAACACCTCGCTCTCCA (SEQ ID NO:8) 7. GCAGGACCGACAGCGCCAGG (SEQ ID NO:9) 8. TCCATCTTTGCCAGCAGGAC (SEQ ID NO:10) 9. GGTCCCTGAGCTCCAGCCTC (SEQ ID NO:11) 10. TTGGCCGTCAGCACCAGGG (SEQ ID NO:12) 11. TTTCTCGCCAGCAGGGCCAG (SEQ ID NO:13) 12. CTCGATCTGCTGGCTCAGGC (SEQ ID NO:14)

Example 2

[0025] Treatment of Cells

[0026] The human cell line WI-26 was grown in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum. The cells were plated in 48-well plates or 6-well plates (Costar, Coming Inc.) to obtain 70-80% confluence. After 24 hours, the cells were washed two times with pre-warmed DMEM and 0.35 ml (for 48-well experiments) or 1 ml (6-well experiments) DMEM containing 5 μg/ml lipofectin (Gibco BRL) or 2.5 μg/ml cytofectin GSV (Glen Research Ltd) and oligonucleotides at 200 nM were added to each well. After 4-5 hours at 37° C. the cells were washed two times with pre-warmed DMEM and 0.35 ml DMEM (48-well plates) or 1 ml DMEM (Swell plates) was added together with ascorbic acid at 20 ug/ml. The cells were incubated for 20 hours prior to analysis of collagen levels.

Example 3

[0027] Protein Analysis

[0028] At the end of the experiment, 150 μl of medium was removed and the amount of secreted type I procollagen determined using an ELISA kit (AmershamPharmacia Ltd) and the results expressed as nanograms of procollagen in the medium/10,000 cells. To correct for cell numbers, plates were washed with pre-warmed PBS, cells treated with trypsin and cell numbers determined using an automated Coulter counter. For Swell experiments, the cells were counted, treated with 1 ml TRI reagent (SIGMA Ltd) and proteins and RNA extracted according to the manufacturers guidelines. The protein pellet was re-suspended in 1% SDS containing protease inhibitors. 30-100 ugs cellular proteins were heated at 100° C. for 5 mins and then lectrophoresed in a 4-12% SDS polyacrylamide gel. Proteins were electrophoretically transferred to nitrocellulose filters and hybridised with an antibody against a synthetic peptide corresponding to human proα1 (1) chain of type I collagen (obtained from Dr Larry Fisher, NIH, USA). The proα1 (1) band was detected using an anti-rabbit secondary antibody coupled to HRP (Biorad Ltd) and developed using ECL (Pierce Ltd). Protein loading was determined by treating the membrane with an antibody to GAPDH (Advanced Immunochemicals). Protein loading was normalised to GAPDH levels using desitometry.

Example 4

[0029] RNA Analysis

[0030] RNA was extracted using TRI reagent and the final pellet was re-suspended in 0.5% SDS. One to three micrograms of total RNA were electrophoresed in a formaldehyde denaturing gel according to standard procedures (Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). RNA was transferred to Hybond-N membranes (Amersham) and hybridised for 24 hours to an alpha1 (1) cDNA probe radiolabeled using a T7 polymerase kit (AmershamPharmacia). Following washing, the filter was exposed to X-ray film and the film developed 4-24 hours later. The autoradiographic images of the alpha1 (1) transcripts (4.8 kb & 5.8 kb) were analysed by densitometric analysis and RNA loading was corrected using the intensity of the GAPDH transcript or the intensity of the 28S rRNA as internal controls.

[0031] Results

[0032] Table I below shows the average percentage (%) collagen inhibition which relates to either collagen levels in the medium or collagen mRNA levels. In the treated cell assay used, there was a very good correlation between percentage collagen inhibition as measured in the medium and percentage inhibition of intracellular collagen mRNA levels. TABLE I AVERAGE % COLLAGEN ASO INHIBITION (SEQ ID NO:3) GGACGACCAGGTTTTCCAGC 50 (SEQ ID NO:4) GCAGCACCAGCAGGGCCAGG 50-80 (SEQ ID NO:5) GCCAGGAGCACCAGGTTCAC 50 (SEQ ID NO:6) CTTCCTCTCCAGCAGGGCCA 50-60 (SEQ ID NO:7) GCCTTGCCGGGCTCTCCAGC 50 (SEQ ID NO:8) CGGGAACACCTCGCTCTCCA 50 (SEQ ID NO:9) GCAGGACCGACAGCGCCAGG 50 (SEQ ID NO:10) TCCATCTTTGCCAGCAGGAC 50 (SEQ ID NO:11) GGTCCCTGAGCTCCAGCCTC 50 (SEQ ID NO:12) TTGGCCGTCAGCACCAGGG 50-80 (SEQ ID NO:13) TTTCTCGCCAGCAGGGCCAG 50-70 (SEQ ID NO:14) CTCGATCTGCTGGCTCAGGC 50-80

[0033]

1 14 1 6728 DNA Homo sapiens CDS (120)..(4511) sig_peptide (120)..(185) 1 agcagacggg agtttctcct cggggtcgga gcaggaggca cgcggagtgt gaggccacgc 60 atgagcggac gctaaccccc tccccagcca caaagagtct acatgtctag ggtctagac 119 atg ttc agc ttt gtg gac ctc cgg ctc ctg ctc ctc tta gcg gcc acc 167 Met Phe Ser Phe Val Asp Leu Arg Leu Leu Leu Leu Leu Ala Ala Thr 1 5 10 15 gcc ctc ctg acg cac ggc caa gag gaa ggc caa gtc gag ggc caa gac 215 Ala Leu Leu Thr His Gly Gln Glu Glu Gly Gln Val Glu Gly Gln Asp 20 25 30 gaa gac atc cca cca atc acc tgc gta cag aac ggc ctc agg tac cat 263 Glu Asp Ile Pro Pro Ile Thr Cys Val Gln Asn Gly Leu Arg Tyr His 35 40 45 gac cga gac gtg tgg aaa ccc gag ccc tgc cgg atc tgc gtc tgc gac 311 Asp Arg Asp Val Trp Lys Pro Glu Pro Cys Arg Ile Cys Val Cys Asp 50 55 60 aac ggc aag gtg ttg tgc gat gac gtg atc tgt gac gag acc aag aac 359 Asn Gly Lys Val Leu Cys Asp Asp Val Ile Cys Asp Glu Thr Lys Asn 65 70 75 80 tgc ccc ggc gcc gaa gtc ccc gag ggc gag tgc tgt ccc gtc tgc ccc 407 Cys Pro Gly Ala Glu Val Pro Glu Gly Glu Cys Cys Pro Val Cys Pro 85 90 95 gac ggc tca gag tca ccc acc gac caa gaa acc acc ggc gtc gag gga 455 Asp Gly Ser Glu Ser Pro Thr Asp Gln Glu Thr Thr Gly Val Glu Gly 100 105 110 ccc aag gga gac act ggc ccc cga ggc cca agg gga ccc gca ggc ccc 503 Pro Lys Gly Asp Thr Gly Pro Arg Gly Pro Arg Gly Pro Ala Gly Pro 115 120 125 cct ggc cga gat ggc atc cct gga cag cct gga ctt ccc gga ccc ccc 551 Pro Gly Arg Asp Gly Ile Pro Gly Gln Pro Gly Leu Pro Gly Pro Pro 130 135 140 gga ccc ccc gga cct ccc gga ccc cct ggc ctc gga gga aac ttt gct 599 Gly Pro Pro Gly Pro Pro Gly Pro Pro Gly Leu Gly Gly Asn Phe Ala 145 150 155 160 ccc cag ctg tct tat ggc tat gat gag aaa tca acc gga gga att tcc 647 Pro Gln Leu Ser Tyr Gly Tyr Asp Glu Lys Ser Thr Gly Gly Ile Ser 165 170 175 gtg cct ggc ccc atg ggt ccc tct ggt cct cgt ggt ctc cct ggc ccc 695 Val Pro Gly Pro Met Gly Pro Ser Gly Pro Arg Gly Leu Pro Gly Pro 180 185 190 cct ggt gca cct ggt ccc caa ggc ttc caa ggt ccc cct ggt gag cct 743 Pro Gly Ala Pro Gly Pro Gln Gly Phe Gln Gly Pro Pro Gly Glu Pro 195 200 205 ggc gag cct gga gct tca ggt ccc atg ggt ccc cga ggt ccc cca ggt 791 Gly Glu Pro Gly Ala Ser Gly Pro Met Gly Pro Arg Gly Pro Pro Gly 210 215 220 ccc cct gga aag aat gga gat gat ggg gaa gct gga aaa cct ggt cgt 839 Pro Pro Gly Lys Asn Gly Asp Asp Gly Glu Ala Gly Lys Pro Gly Arg 225 230 235 240 cct ggt gag cgt ggg cct cct ggg cct cag ggt gct cga gga ttg ccc 887 Pro Gly Glu Arg Gly Pro Pro Gly Pro Gln Gly Ala Arg Gly Leu Pro 245 250 255 gga aca gct ggc ctc cct gga atg aag gga cac aga ggt ttc agt ggt 935 Gly Thr Ala Gly Leu Pro Gly Met Lys Gly His Arg Gly Phe Ser Gly 260 265 270 ttg gat ggt gcc aag gga gat gct ggt cct gct ggt cct aag ggt gag 983 Leu Asp Gly Ala Lys Gly Asp Ala Gly Pro Ala Gly Pro Lys Gly Glu 275 280 285 cct ggc agc cct ggt gaa aat gga gct cct ggt cag atg ggc ccc cgt 1031 Pro Gly Ser Pro Gly Glu Asn Gly Ala Pro Gly Gln Met Gly Pro Arg 290 295 300 ggc ctg cct ggt gag aga ggt cgc cct gga gcc cct ggc cct gct ggt 1079 Gly Leu Pro Gly Glu Arg Gly Arg Pro Gly Ala Pro Gly Pro Ala Gly 305 310 315 320 gct cgt gga aat gat ggt gct act ggt gct gcc ggg ccc cct ggt ccc 1127 Ala Arg Gly Asn Asp Gly Ala Thr Gly Ala Ala Gly Pro Pro Gly Pro 325 330 335 acc ggc ccc gct ggt cct cct ggc ttc cct ggt gct gtt ggt gct aag 1175 Thr Gly Pro Ala Gly Pro Pro Gly Phe Pro Gly Ala Val Gly Ala Lys 340 345 350 ggt gaa gct ggt ccc caa ggg ccc cga ggc tct gaa ggt ccc cag ggt 1223 Gly Glu Ala Gly Pro Gln Gly Pro Arg Gly Ser Glu Gly Pro Gln Gly 355 360 365 gtg cgt ggt gag cct ggc ccc cct ggc cct gct ggt gct gct ggc cct 1271 Val Arg Gly Glu Pro Gly Pro Pro Gly Pro Ala Gly Ala Ala Gly Pro 370 375 380 gct gga aac cct ggt gct gat gga cag cct ggt gct aaa ggt gcc aat 1319 Ala Gly Asn Pro Gly Ala Asp Gly Gln Pro Gly Ala Lys Gly Ala Asn 385 390 395 400 ggt gct cct ggt att gct ggt gct cct ggc ttc cct ggt gcc cga ggc 1367 Gly Ala Pro Gly Ile Ala Gly Ala Pro Gly Phe Pro Gly Ala Arg Gly 405 410 415 ccc tct gga ccc cag ggc ccc ggc ggc cct cct ggt ccc aag ggt aac 1415 Pro Ser Gly Pro Gln Gly Pro Gly Gly Pro Pro Gly Pro Lys Gly Asn 420 425 430 agc ggt gaa cct ggt gct cct ggc agc aaa gga gac act ggt gct aag 1463 Ser Gly Glu Pro Gly Ala Pro Gly Ser Lys Gly Asp Thr Gly Ala Lys 435 440 445 gga gag cct ggc cct gtt ggt gtt caa gga ccc cct ggc cct gct gga 1511 Gly Glu Pro Gly Pro Val Gly Val Gln Gly Pro Pro Gly Pro Ala Gly 450 455 460 gag gaa gga aag cga gga gct cga ggt gaa ccc gga ccc act ggc ctg 1559 Glu Glu Gly Lys Arg Gly Ala Arg Gly Glu Pro Gly Pro Thr Gly Leu 465 470 475 480 ccc gga ccc cct ggc gag cgt ggt gga cct ggt agc cgt ggt ttc cct 1607 Pro Gly Pro Pro Gly Glu Arg Gly Gly Pro Gly Ser Arg Gly Phe Pro 485 490 495 ggc gca gat ggt gtt gct ggt ccc aag ggt ccc gct ggt gaa cgt ggt 1655 Gly Ala Asp Gly Val Ala Gly Pro Lys Gly Pro Ala Gly Glu Arg Gly 500 505 510 tct cct ggc ccc gct ggc ccc aaa gga tct cct ggt gaa gct ggt cgt 1703 Ser Pro Gly Pro Ala Gly Pro Lys Gly Ser Pro Gly Glu Ala Gly Arg 515 520 525 ccc ggt gaa gct ggt ctg cct ggt gcc aag ggt ctg act gga agc cct 1751 Pro Gly Glu Ala Gly Leu Pro Gly Ala Lys Gly Leu Thr Gly Ser Pro 530 535 540 ggc agc cct ggt cct gat ggc aaa act ggc ccc cct ggt ccc gcc ggt 1799 Gly Ser Pro Gly Pro Asp Gly Lys Thr Gly Pro Pro Gly Pro Ala Gly 545 550 555 560 caa gat ggt cgc ccc gga ccc cca ggc cca cct ggt gcc cgt ggt cag 1847 Gln Asp Gly Arg Pro Gly Pro Pro Gly Pro Pro Gly Ala Arg Gly Gln 565 570 575 gct ggt gtg atg gga ttc cct gga cct aaa ggt gct gct gga gag ccc 1895 Ala Gly Val Met Gly Phe Pro Gly Pro Lys Gly Ala Ala Gly Glu Pro 580 585 590 ggc aag gct gga gag cga ggt gtt ccc gga ccc cct ggc gct gtc ggt 1943 Gly Lys Ala Gly Glu Arg Gly Val Pro Gly Pro Pro Gly Ala Val Gly 595 600 605 cct gct ggc aaa gat gga gag gct gga gct cag gga ccc cct ggc cct 1991 Pro Ala Gly Lys Asp Gly Glu Ala Gly Ala Gln Gly Pro Pro Gly Pro 610 615 620 gct ggt ccc gct ggc gag aga ggt gaa caa ggc cct gct ggc tcc ccc 2039 Ala Gly Pro Ala Gly Glu Arg Gly Glu Gln Gly Pro Ala Gly Ser Pro 625 630 635 640 gga ttc cag ggt ctc cct ggt cct gct ggt cct cca ggt gaa gca ggc 2087 Gly Phe Gln Gly Leu Pro Gly Pro Ala Gly Pro Pro Gly Glu Ala Gly 645 650 655 aaa cct ggt gaa cag ggt gtt cct gga gac ctt ggc gcc cct ggc ccc 2135 Lys Pro Gly Glu Gln Gly Val Pro Gly Asp Leu Gly Ala Pro Gly Pro 660 665 670 tct gga gca aga ggc gag aga ggt ttc cct ggc gag cgt ggt gtg caa 2183 Ser Gly Ala Arg Gly Glu Arg Gly Phe Pro Gly Glu Arg Gly Val Gln 675 680 685 ggt ccc cct ggt cct gct gga ccc cga ggg gcc aac ggt gct ccc ggc 2231 Gly Pro Pro Gly Pro Ala Gly Pro Arg Gly Ala Asn Gly Ala Pro Gly 690 695 700 aac gat ggt gct aag ggt gat gct ggt gcc cct gga gct ccc ggt agc 2279 Asn Asp Gly Ala Lys Gly Asp Ala Gly Ala Pro Gly Ala Pro Gly Ser 705 710 715 720 cag ggc gcc cct ggc ctt cag gga atg cct ggt gaa cgt ggt gca gct 2327 Gln Gly Ala Pro Gly Leu Gln Gly Met Pro Gly Glu Arg Gly Ala Ala 725 730 735 ggt ctt cca ggg cct aag ggt gac aga ggt gat gct ggt ccc aaa ggt 2375 Gly Leu Pro Gly Pro Lys Gly Asp Arg Gly Asp Ala Gly Pro Lys Gly 740 745 750 gct gat ggc tct cct ggc aaa gat ggc gtc cgt ggt ctg acc ggc ccc 2423 Ala Asp Gly Ser Pro Gly Lys Asp Gly Val Arg Gly Leu Thr Gly Pro 755 760 765 att ggt cct cct ggc cct gct ggt gcc cct ggt gac aag ggt gaa agt 2471 Ile Gly Pro Pro Gly Pro Ala Gly Ala Pro Gly Asp Lys Gly Glu Ser 770 775 780 ggt ccc agc ggc cct gct ggt ccc act gga gct cgt ggt gcc ccc gga 2519 Gly Pro Ser Gly Pro Ala Gly Pro Thr Gly Ala Arg Gly Ala Pro Gly 785 790 795 800 gac cgt ggt gag cct ggt ccc ccc ggc cct gct ggc ttt gct ggc ccc 2567 Asp Arg Gly Glu Pro Gly Pro Pro Gly Pro Ala Gly Phe Ala Gly Pro 805 810 815 cct ggt gct gac ggc caa cct ggt gct aaa ggc gaa cct ggt gat gct 2615 Pro Gly Ala Asp Gly Gln Pro Gly Ala Lys Gly Glu Pro Gly Asp Ala 820 825 830 ggt gcc aaa ggc gat gct ggt ccc cct ggg cct gcc gga ccc gct gga 2663 Gly Ala Lys Gly Asp Ala Gly Pro Pro Gly Pro Ala Gly Pro Ala Gly 835 840 845 ccc cct ggc ccc att ggt aat gtt ggt gct cct gga gcc aaa ggt gct 2711 Pro Pro Gly Pro Ile Gly Asn Val Gly Ala Pro Gly Ala Lys Gly Ala 850 855 860 cgc ggc agc gct ggt ccc cct ggt gct act ggt ttc cct ggt gct gct 2759 Arg Gly Ser Ala Gly Pro Pro Gly Ala Thr Gly Phe Pro Gly Ala Ala 865 870 875 880 ggc cga gtc ggt cct cct ggc ccc tct gga aat gct gga ccc cct ggc 2807 Gly Arg Val Gly Pro Pro Gly Pro Ser Gly Asn Ala Gly Pro Pro Gly 885 890 895 cct cct ggt cct gct ggc aaa gaa ggc ggc aaa ggt ccc cgt ggt gag 2855 Pro Pro Gly Pro Ala Gly Lys Glu Gly Gly Lys Gly Pro Arg Gly Glu 900 905 910 act ggc cct gct gga cgt cct ggt gaa gtt ggt ccc cct ggt ccc cct 2903 Thr Gly Pro Ala Gly Arg Pro Gly Glu Val Gly Pro Pro Gly Pro Pro 915 920 925 ggc cct gct ggc gag aaa gga tcc cct ggt gct gat ggt cct gct ggt 2951 Gly Pro Ala Gly Glu Lys Gly Ser Pro Gly Ala Asp Gly Pro Ala Gly 930 935 940 gct cct ggt act ccc ggg cct caa ggt att gct gga cag cgt ggt gtg 2999 Ala Pro Gly Thr Pro Gly Pro Gln Gly Ile Ala Gly Gln Arg Gly Val 945 950 955 960 gtc ggc ctg cct ggt cag aga gga gag aga ggc ttc cct ggt ctt cct 3047 Val Gly Leu Pro Gly Gln Arg Gly Glu Arg Gly Phe Pro Gly Leu Pro 965 970 975 ggc ccc tct ggt gaa cct ggc aaa caa ggt ccc tct gga gca agt ggt 3095 Gly Pro Ser Gly Glu Pro Gly Lys Gln Gly Pro Ser Gly Ala Ser Gly 980 985 990 gaa cgt ggt ccc ccc ggt ccc atg ggc ccc cct gga ttg gct gga ccc 3143 Glu Arg Gly Pro Pro Gly Pro Met Gly Pro Pro Gly Leu Ala Gly Pro 995 1000 1005 cct ggt gaa tct gga cgt gag ggg gct cct gct gcc gaa ggt tcc cct 3191 Pro Gly Glu Ser Gly Arg Glu Gly Ala Pro Ala Ala Glu Gly Ser Pro 1010 1015 1020 gga cga gac ggt tct cct ggc gcc aag ggt gac cgt ggt gag acc ggc 3239 Gly Arg Asp Gly Ser Pro Gly Ala Lys Gly Asp Arg Gly Glu Thr Gly 1025 1030 1035 1040 ccc gct gga ccc cct ggt gct cct ggt gct cct ggt gcc cct ggc ccc 3287 Pro Ala Gly Pro Pro Gly Ala Pro Gly Ala Pro Gly Ala Pro Gly Pro 1045 1050 1055 gtt ggc cct gct ggc aag agt ggt gat cgt ggt gag act ggt cct gct 3335 Val Gly Pro Ala Gly Lys Ser Gly Asp Arg Gly Glu Thr Gly Pro Ala 1060 1065 1070 ggt ccc gcc ggt ccc gtc ggc ccc gtc ggc gcc cgt ggc ccc gcc gga 3383 Gly Pro Ala Gly Pro Val Gly Pro Val Gly Ala Arg Gly Pro Ala Gly 1075 1080 1085 ccc caa ggc ccc cgt ggt gac aag ggt gag aca ggc gaa cag ggc gac 3431 Pro Gln Gly Pro Arg Gly Asp Lys Gly Glu Thr Gly Glu Gln Gly Asp 1090 1095 1100 aga ggc ata aag ggt cac cgt ggc ttc tct ggc ctc cag ggt ccc cct 3479 Arg Gly Ile Lys Gly His Arg Gly Phe Ser Gly Leu Gln Gly Pro Pro 1105 1110 1115 1120 ggc cct cct ggc tct cct ggt gaa caa ggt ccc tct gga gcc tct ggt 3527 Gly Pro Pro Gly Ser Pro Gly Glu Gln Gly Pro Ser Gly Ala Ser Gly 1125 1130 1135 cct gct ggt ccc cga ggt ccc cct ggc tct gct ggt gct cct ggc aaa 3575 Pro Ala Gly Pro Arg Gly Pro Pro Gly Ser Ala Gly Ala Pro Gly Lys 1140 1145 1150 gat gga ctc aac ggt ctc cct ggc ccc att ggg ccc cct ggt cct cgc 3623 Asp Gly Leu Asn Gly Leu Pro Gly Pro Ile Gly Pro Pro Gly Pro Arg 1155 1160 1165 ggt cgc act ggt gat gct ggt cct gtt ggt ccc ccc ggc cct cct gga 3671 Gly Arg Thr Gly Asp Ala Gly Pro Val Gly Pro Pro Gly Pro Pro Gly 1170 1175 1180 cct cct ggt ccc cct ggt cct ccc agc gct ggt ttc gac ttc agc ttc 3719 Pro Pro Gly Pro Pro Gly Pro Pro Ser Ala Gly Phe Asp Phe Ser Phe 1185 1190 1195 1200 ctg ccc cag cca cct caa gag aag gct cac gat ggt ggc cgc tac tac 3767 Leu Pro Gln Pro Pro Gln Glu Lys Ala His Asp Gly Gly Arg Tyr Tyr 1205 1210 1215 cgg gct gat gat gcc aat gtg gtt cgt gac cgt gac ctc gag gtg gac 3815 Arg Ala Asp Asp Ala Asn Val Val Arg Asp Arg Asp Leu Glu Val Asp 1220 1225 1230 acc acc ctc aag agc ctg agc cag cag atc gag aac atc cgg agc cca 3863 Thr Thr Leu Lys Ser Leu Ser Gln Gln Ile Glu Asn Ile Arg Ser Pro 1235 1240 1245 gag gga agc cgc aag aac ccc gcc cgc acc tgc cgt gac ctc aag atg 3911 Glu Gly Ser Arg Lys Asn Pro Ala Arg Thr Cys Arg Asp Leu Lys Met 1250 1255 1260 tgc cac tct gac tgg aag agt gga gag tac tgg att gac ccc aac caa 3959 Cys His Ser Asp Trp Lys Ser Gly Glu Tyr Trp Ile Asp Pro Asn Gln 1265 1270 1275 1280 ggc tgc aac ctg gat gcc atc aaa gtc ttc tgc aac atg gag act ggt 4007 Gly Cys Asn Leu Asp Ala Ile Lys Val Phe Cys Asn Met Glu Thr Gly 1285 1290 1295 gag acc tgc gtg tac ccc act cag ccc agt gtg gcc cag aag aac tgg 4055 Glu Thr Cys Val Tyr Pro Thr Gln Pro Ser Val Ala Gln Lys Asn Trp 1300 1305 1310 tac atc agc aag aac ccc aag gac aag agg cat gtc tgg ttc ggc gag 4103 Tyr Ile Ser Lys Asn Pro Lys Asp Lys Arg His Val Trp Phe Gly Glu 1315 1320 1325 agc atg acc gat gga ttc cag ttc gag tat ggc ggc cag ggc tcc gac 4151 Ser Met Thr Asp Gly Phe Gln Phe Glu Tyr Gly Gly Gln Gly Ser Asp 1330 1335 1340 cct gcc gat gtg gcc atc cag ctg acc ttc ctg cgc ctg atg tcc acc 4199 Pro Ala Asp Val Ala Ile Gln Leu Thr Phe Leu Arg Leu Met Ser Thr 1345 1350 1355 1360 gag gcc tcc cag aac atc acc tac cac tgc aag aac agc gtg gcc tac 4247 Glu Ala Ser Gln Asn Ile Thr Tyr His Cys Lys Asn Ser Val Ala Tyr 1365 1370 1375 atg gac cag cag act ggc aac ctc aag aag gcc ctg ctc ctc aag ggc 4295 Met Asp Gln Gln Thr Gly Asn Leu Lys Lys Ala Leu Leu Leu Lys Gly 1380 1385 1390 tcc aac gag atc gag atc cgc gcc gag ggc aac agc cgc ttc acc tac 4343 Ser Asn Glu Ile Glu Ile Arg Ala Glu Gly Asn Ser Arg Phe Thr Tyr 1395 1400 1405 agc gtc act gtc gat ggc tgc acg agt cac acc gga gcc tgg ggc aag 4391 Ser Val Thr Val Asp Gly Cys Thr Ser His Thr Gly Ala Trp Gly Lys 1410 1415 1420 aca gtg att gaa tac aaa acc acc aag tcc tcc cgc ctg ccc atc atc 4439 Thr Val Ile Glu Tyr Lys Thr Thr Lys Ser Ser Arg Leu Pro Ile Ile 1425 1430 1435 1440 gat gtg gcc ccc ttg gac gtt ggt gcc cca gac cag gaa ttc ggc ttc 4487 Asp Val Ala Pro Leu Asp Val Gly Ala Pro Asp Gln Glu Phe Gly Phe 1445 1450 1455 gac gtt ggc cct gtc tgc ttc ctg taaactccct ccatcccaac ctggctccct 4541 Asp Val Gly Pro Val Cys Phe Leu 1460 cccacccaac caactttccc cccaacccgg aaacagacaa gcaacccaaa ctgaaccccc 4601 ccaaaagcca aaaaatggga gacaatttca catggacttt ggaaaatatt tttttccttt 4661 gcattcatct ctcaaactta gtttttatct ttgaccaacc gaacatgacc aaaaaccaaa 4721 agtgcattca accttaccaa aaaaaaaaaa aaaaaaaaaa gaataaataa ataagttttt 4781 aaaaaaggaa gcttggtcca cttgcttgaa gacccatgcg ggggtaagtc cctttctgcc 4841 cgttgggtta tgaaacccca atgctgccct ttctgctcct ttctccacac cccccttggc 4901 ctcccctcca ctccttccca aatctgtctc cccagaagac acaggaaaca atgtattgtc 4961 tgcccagcaa tcaaaggcaa tgctcaaaca cccaagtggc ccccaccctc agcccgctcc 5021 tgcccgccca gcacccccag gccctgggga cctggggttc tcagactgcc aaagaagcct 5081 tgccatctgg cgctcccatg gctcttgcaa catctcccct tcgtttttga gggggtcatg 5141 ccgggggagc caccagcccc tcactgggtt cggaggagag tcaggaaggg ccacgacaaa 5201 gcagaaacat cggatttggg gaacgcgtgt catcccttgt gccgcaggct gggcgggaga 5261 gactgttctg ttctgttcct tgtgtaactg tgttgctgaa agactacctc gttcttgtct 5321 tgatgtgtca ccggggcaac tgcctggggg cggggatggg ggcagggtgg aagcggctcc 5381 ccatttttat accaaaggtg ctacatctat gtgatgggtg gggtggggag ggaatcactg 5441 gtgctataga aattgagatg cccccccagg ccagcaaatg ttcctttttg ttcaaagtct 5501 atttttattc cttgatattt tttctttctt tttttttttt tttgtggatg gggacttgtg 5561 aatttttcta aaggtgctat ttaacatggg aggagagcgt gtgcgctcca gcccagcccg 5621 ctgctcactt tccaccctct ctccacctgc ctctggcttc tcaggcctct gctctccgac 5681 ctctctcctc tgaaaccctc ctccacagct gcagcccatc ctcccggctc cctcctagtc 5741 tgtcctgcgt cctctgtccc cgggtttcag agacaacttc ccaaagcaca aagcagtttt 5801 tccctagggg tgggaggaag caaaagactc tgtacctatt ttgtatgtgt ataataattt 5861 gagatgtttt taattatttt gattgctgga ataaagcatg tggaaatgac ccaaacataa 5921 tccgcagtgg cctcctaatt tccttctttg gagttggggg aggggtagac atggggaagg 5981 ggccttgggg tgatgggctt gccttccatt cctgcccttt ccctccccac tattctcttc 6041 tagatccctc cataacccca ctcccctttc tctcaccctt cttataccgc aaacctttct 6101 acttcctctt tcattttcta ttcttgcaat ttccttgcac cttttccaaa tcctcttctc 6161 ccctgcaata ccatacaggc aatccacgtg cacaacacac acacacactc ttcacatctg 6221 gggttgtcca aacctcatac ccactcccct tcaagcccat ccactctcca ccccctggat 6281 gccctgcact tggtggcggt gggatgctca tggatactgg gagggtgagg ggagtggaac 6341 ccgtgaggag gacctggggg cctctccttg aactgacatg aagggtcatc tggcctctgc 6401 tcccttctca cccacgctga cctcctgccg aaggagcaac gcaacaggag aggggtctgc 6461 tgagcctggc gagggtctgg gagggaccag gaggaaggcg tgctccctgc tcgctgtcct 6521 ggccctgggg gagtgaggga gacagacacc tgggagagct gtggggaagg cactcgcacc 6581 gtgctcttgg gaaggaagga gacctggccc tgctcaccac ggactgggtg cctcgacctc 6641 ctgaatcccc agaacacaac ccccctgggc tggggtggtc tggggaacca tcgtgccccc 6701 gcctcccgcc tactcctttt taagctt 6728 2 1464 PRT Homo sapiens 2 Met Phe Ser Phe Val Asp Leu Arg Leu Leu Leu Leu Leu Ala Ala Thr 1 5 10 15 Ala Leu Leu Thr His Gly Gln Glu Glu Gly Gln Val Glu Gly Gln Asp 20 25 30 Glu Asp Ile Pro Pro Ile Thr Cys Val Gln Asn Gly Leu Arg Tyr His 35 40 45 Asp Arg Asp Val Trp Lys Pro Glu Pro Cys Arg Ile Cys Val Cys Asp 50 55 60 Asn Gly Lys Val Leu Cys Asp Asp Val Ile Cys Asp Glu Thr Lys Asn 65 70 75 80 Cys Pro Gly Ala Glu Val Pro Glu Gly Glu Cys Cys Pro Val Cys Pro 85 90 95 Asp Gly Ser Glu Ser Pro Thr Asp Gln Glu Thr Thr Gly Val Glu Gly 100 105 110 Pro Lys Gly Asp Thr Gly Pro Arg Gly Pro Arg Gly Pro Ala Gly Pro 115 120 125 Pro Gly Arg Asp Gly Ile Pro Gly Gln Pro Gly Leu Pro Gly Pro Pro 130 135 140 Gly Pro Pro Gly Pro Pro Gly Pro Pro Gly Leu Gly Gly Asn Phe Ala 145 150 155 160 Pro Gln Leu Ser Tyr Gly Tyr Asp Glu Lys Ser Thr Gly Gly Ile Ser 165 170 175 Val Pro Gly Pro Met Gly Pro Ser Gly Pro Arg Gly Leu Pro Gly Pro 180 185 190 Pro Gly Ala Pro Gly Pro Gln Gly Phe Gln Gly Pro Pro Gly Glu Pro 195 200 205 Gly Glu Pro Gly Ala Ser Gly Pro Met Gly Pro Arg Gly Pro Pro Gly 210 215 220 Pro Pro Gly Lys Asn Gly Asp Asp Gly Glu Ala Gly Lys Pro Gly Arg 225 230 235 240 Pro Gly Glu Arg Gly Pro Pro Gly Pro Gln Gly Ala Arg Gly Leu Pro 245 250 255 Gly Thr Ala Gly Leu Pro Gly Met Lys Gly His Arg Gly Phe Ser Gly 260 265 270 Leu Asp Gly Ala Lys Gly Asp Ala Gly Pro Ala Gly Pro Lys Gly Glu 275 280 285 Pro Gly Ser Pro Gly Glu Asn Gly Ala Pro Gly Gln Met Gly Pro Arg 290 295 300 Gly Leu Pro Gly Glu Arg Gly Arg Pro Gly Ala Pro Gly Pro Ala Gly 305 310 315 320 Ala Arg Gly Asn Asp Gly Ala Thr Gly Ala Ala Gly Pro Pro Gly Pro 325 330 335 Thr Gly Pro Ala Gly Pro Pro Gly Phe Pro Gly Ala Val Gly Ala Lys 340 345 350 Gly Glu Ala Gly Pro Gln Gly Pro Arg Gly Ser Glu Gly Pro Gln Gly 355 360 365 Val Arg Gly Glu Pro Gly Pro Pro Gly Pro Ala Gly Ala Ala Gly Pro 370 375 380 Ala Gly Asn Pro Gly Ala Asp Gly Gln Pro Gly Ala Lys Gly Ala Asn 385 390 395 400 Gly Ala Pro Gly Ile Ala Gly Ala Pro Gly Phe Pro Gly Ala Arg Gly 405 410 415 Pro Ser Gly Pro Gln Gly Pro Gly Gly Pro Pro Gly Pro Lys Gly Asn 420 425 430 Ser Gly Glu Pro Gly Ala Pro Gly Ser Lys Gly Asp Thr Gly Ala Lys 435 440 445 Gly Glu Pro Gly Pro Val Gly Val Gln Gly Pro Pro Gly Pro Ala Gly 450 455 460 Glu Glu Gly Lys Arg Gly Ala Arg Gly Glu Pro Gly Pro Thr Gly Leu 465 470 475 480 Pro Gly Pro Pro Gly Glu Arg Gly Gly Pro Gly Ser Arg Gly Phe Pro 485 490 495 Gly Ala Asp Gly Val Ala Gly Pro Lys Gly Pro Ala Gly Glu Arg Gly 500 505 510 Ser Pro Gly Pro Ala Gly Pro Lys Gly Ser Pro Gly Glu Ala Gly Arg 515 520 525 Pro Gly Glu Ala Gly Leu Pro Gly Ala Lys Gly Leu Thr Gly Ser Pro 530 535 540 Gly Ser Pro Gly Pro Asp Gly Lys Thr Gly Pro Pro Gly Pro Ala Gly 545 550 555 560 Gln Asp Gly Arg Pro Gly Pro Pro Gly Pro Pro Gly Ala Arg Gly Gln 565 570 575 Ala Gly Val Met Gly Phe Pro Gly Pro Lys Gly Ala Ala Gly Glu Pro 580 585 590 Gly Lys Ala Gly Glu Arg Gly Val Pro Gly Pro Pro Gly Ala Val Gly 595 600 605 Pro Ala Gly Lys Asp Gly Glu Ala Gly Ala Gln Gly Pro Pro Gly Pro 610 615 620 Ala Gly Pro Ala Gly Glu Arg Gly Glu Gln Gly Pro Ala Gly Ser Pro 625 630 635 640 Gly Phe Gln Gly Leu Pro Gly Pro Ala Gly Pro Pro Gly Glu Ala Gly 645 650 655 Lys Pro Gly Glu Gln Gly Val Pro Gly Asp Leu Gly Ala Pro Gly Pro 660 665 670 Ser Gly Ala Arg Gly Glu Arg Gly Phe Pro Gly Glu Arg Gly Val Gln 675 680 685 Gly Pro Pro Gly Pro Ala Gly Pro Arg Gly Ala Asn Gly Ala Pro Gly 690 695 700 Asn Asp Gly Ala Lys Gly Asp Ala Gly Ala Pro Gly Ala Pro Gly Ser 705 710 715 720 Gln Gly Ala Pro Gly Leu Gln Gly Met Pro Gly Glu Arg Gly Ala Ala 725 730 735 Gly Leu Pro Gly Pro Lys Gly Asp Arg Gly Asp Ala Gly Pro Lys Gly 740 745 750 Ala Asp Gly Ser Pro Gly Lys Asp Gly Val Arg Gly Leu Thr Gly Pro 755 760 765 Ile Gly Pro Pro Gly Pro Ala Gly Ala Pro Gly Asp Lys Gly Glu Ser 770 775 780 Gly Pro Ser Gly Pro Ala Gly Pro Thr Gly Ala Arg Gly Ala Pro Gly 785 790 795 800 Asp Arg Gly Glu Pro Gly Pro Pro Gly Pro Ala Gly Phe Ala Gly Pro 805 810 815 Pro Gly Ala Asp Gly Gln Pro Gly Ala Lys Gly Glu Pro Gly Asp Ala 820 825 830 Gly Ala Lys Gly Asp Ala Gly Pro Pro Gly Pro Ala Gly Pro Ala Gly 835 840 845 Pro Pro Gly Pro Ile Gly Asn Val Gly Ala Pro Gly Ala Lys Gly Ala 850 855 860 Arg Gly Ser Ala Gly Pro Pro Gly Ala Thr Gly Phe Pro Gly Ala Ala 865 870 875 880 Gly Arg Val Gly Pro Pro Gly Pro Ser Gly Asn Ala Gly Pro Pro Gly 885 890 895 Pro Pro Gly Pro Ala Gly Lys Glu Gly Gly Lys Gly Pro Arg Gly Glu 900 905 910 Thr Gly Pro Ala Gly Arg Pro Gly Glu Val Gly Pro Pro Gly Pro Pro 915 920 925 Gly Pro Ala Gly Glu Lys Gly Ser Pro Gly Ala Asp Gly Pro Ala Gly 930 935 940 Ala Pro Gly Thr Pro Gly Pro Gln Gly Ile Ala Gly Gln Arg Gly Val 945 950 955 960 Val Gly Leu Pro Gly Gln Arg Gly Glu Arg Gly Phe Pro Gly Leu Pro 965 970 975 Gly Pro Ser Gly Glu Pro Gly Lys Gln Gly Pro Ser Gly Ala Ser Gly 980 985 990 Glu Arg Gly Pro Pro Gly Pro Met Gly Pro Pro Gly Leu Ala Gly Pro 995 1000 1005 Pro Gly Glu Ser Gly Arg Glu Gly Ala Pro Ala Ala Glu Gly Ser Pro 1010 1015 1020 Gly Arg Asp Gly Ser Pro Gly Ala Lys Gly Asp Arg Gly Glu Thr Gly 1025 1030 1035 1040 Pro Ala Gly Pro Pro Gly Ala Pro Gly Ala Pro Gly Ala Pro Gly Pro 1045 1050 1055 Val Gly Pro Ala Gly Lys Ser Gly Asp Arg Gly Glu Thr Gly Pro Ala 1060 1065 1070 Gly Pro Ala Gly Pro Val Gly Pro Val Gly Ala Arg Gly Pro Ala Gly 1075 1080 1085 Pro Gln Gly Pro Arg Gly Asp Lys Gly Glu Thr Gly Glu Gln Gly Asp 1090 1095 1100 Arg Gly Ile Lys Gly His Arg Gly Phe Ser Gly Leu Gln Gly Pro Pro 1105 1110 1115 1120 Gly Pro Pro Gly Ser Pro Gly Glu Gln Gly Pro Ser Gly Ala Ser Gly 1125 1130 1135 Pro Ala Gly Pro Arg Gly Pro Pro Gly Ser Ala Gly Ala Pro Gly Lys 1140 1145 1150 Asp Gly Leu Asn Gly Leu Pro Gly Pro Ile Gly Pro Pro Gly Pro Arg 1155 1160 1165 Gly Arg Thr Gly Asp Ala Gly Pro Val Gly Pro Pro Gly Pro Pro Gly 1170 1175 1180 Pro Pro Gly Pro Pro Gly Pro Pro Ser Ala Gly Phe Asp Phe Ser Phe 1185 1190 1195 1200 Leu Pro Gln Pro Pro Gln Glu Lys Ala His Asp Gly Gly Arg Tyr Tyr 1205 1210 1215 Arg Ala Asp Asp Ala Asn Val Val Arg Asp Arg Asp Leu Glu Val Asp 1220 1225 1230 Thr Thr Leu Lys Ser Leu Ser Gln Gln Ile Glu Asn Ile Arg Ser Pro 1235 1240 1245 Glu Gly Ser Arg Lys Asn Pro Ala Arg Thr Cys Arg Asp Leu Lys Met 1250 1255 1260 Cys His Ser Asp Trp Lys Ser Gly Glu Tyr Trp Ile Asp Pro Asn Gln 1265 1270 1275 1280 Gly Cys Asn Leu Asp Ala Ile Lys Val Phe Cys Asn Met Glu Thr Gly 1285 1290 1295 Glu Thr Cys Val Tyr Pro Thr Gln Pro Ser Val Ala Gln Lys Asn Trp 1300 1305 1310 Tyr Ile Ser Lys Asn Pro Lys Asp Lys Arg His Val Trp Phe Gly Glu 1315 1320 1325 Ser Met Thr Asp Gly Phe Gln Phe Glu Tyr Gly Gly Gln Gly Ser Asp 1330 1335 1340 Pro Ala Asp Val Ala Ile Gln Leu Thr Phe Leu Arg Leu Met Ser Thr 1345 1350 1355 1360 Glu Ala Ser Gln Asn Ile Thr Tyr His Cys Lys Asn Ser Val Ala Tyr 1365 1370 1375 Met Asp Gln Gln Thr Gly Asn Leu Lys Lys Ala Leu Leu Leu Lys Gly 1380 1385 1390 Ser Asn Glu Ile Glu Ile Arg Ala Glu Gly Asn Ser Arg Phe Thr Tyr 1395 1400 1405 Ser Val Thr Val Asp Gly Cys Thr Ser His Thr Gly Ala Trp Gly Lys 1410 1415 1420 Thr Val Ile Glu Tyr Lys Thr Thr Lys Ser Ser Arg Leu Pro Ile Ile 1425 1430 1435 1440 Asp Val Ala Pro Leu Asp Val Gly Ala Pro Asp Gln Glu Phe Gly Phe 1445 1450 1455 Asp Val Gly Pro Val Cys Phe Leu 1460 3 20 DNA Homo sapiens Antisense oligonucleotide 3 ggacgaccag gttttccagc 20 4 20 DNA Homo sapiens Antisense oligonucleotide 4 gcagcaccag cagggccagg 20 5 20 DNA Homo sapiens Antisense oligonucleotide 5 gccaggagca ccaggttcac 20 6 20 DNA Homo sapiens Antisense oligonucleotide 6 cttcctctcc agcagggcca 20 7 20 DNA Homo sapiens Antisense oligonucleotide 7 gccttgccgg gctctccagc 20 8 20 DNA Homo sapiens Antisense oligonucleotide 8 cgggaacacc tcgctctcca 20 9 20 DNA Homo sapiens Antisense oligonucleotide 9 gcaggaccga cagcgccagg 20 10 20 DNA Homo sapiens Antisense oligonucleotide 10 tccatctttg ccagcaggac 20 11 20 DNA Homo sapiens Antisense oligonucleotide 11 ggtccctgag ctccagcctc 20 12 19 DNA Homo sapiens Antisense oligonucleotide 12 ttggccgtca gcaccaggg 19 13 20 DNA Homo sapiens Antisense oligonucleotide 13 tttctcgcca gcagggccag 20 14 20 DNA Homo sapiens Antisense oligonucleotide 14 ctcgatctgc tggctcaggc 20 

1. An antisense DNA oligonucleotide comprising from 18 to 25 nucleotides which is complementary to a nucleotide sequence from position 750 to position 3900 inclusive of SEQ ID NO:1, wherein SEQ ID NO:1 comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence according to SEQ ID NO:2, the oligonucleotide being capable of inhibiting expression of the polypeptide in a cell that expresses it.
 2. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 750 to position 900 inclusive of SEQ ID NO:1.
 3. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 1200 to position 1300 inclusive of SEQ ID NO:1.
 4. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 1400 to position 1500 inclusive of SEQ ID NO:1.
 5. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 1450 to position 1550 inclusive of SEQ ID NO:1.
 6. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 1850 to position 2000 inclusive of SEQ ID NO:1.
 7. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 2500 to position 2600 inclusive of SEQ ID NO:1.
 8. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 2850 to position 2950 inclusive of SEQ ID NO:1.
 9. An oligonucleotide according to claim 1 which is complementary to a nucleotide sequence from position 3800 to position 3900 inclusive of SEQ ID NO:1.
 10. An oligonucleotide according to claim 1 which is selected from SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 and SEQ ID NO:14.
 11. A pharmaceutical composition comprising an oligonucleotide as defined in any one of claims 1 to 10 in association with a pharmaceutically acceptable adjuvant, diluent or carrier.
 12. A process for the preparation of a pharmaceutical composition as claimed in claim 11, which comprises mixing an oligonucleotide as defined in any one of claims 1 to 10 with a pharmaceutically acceptable adjuvant, diluent or carrier.
 13. An oligonucleotide according to any one of claims 1 to 10 for use in therapy.
 14. Use of an oligonucleotide according to any one of claims 1 to 10 in the manufacture of a medicament for use in therapy.
 15. A method of treating, or reducing the risk of, a collagen disorder in a patient suffering from, or at risk of, the disorder, which comprises administering to the patient a therapeutically effective amount of an oligonucleotide as defined in any one of claims 1 to 10 or a pharmaceutical composition as defined in claim
 11. 